Expanding the chemical scope of RNA : methyltransferases to site-specific alkynylation of RNA for click labeling

Autoren

Yuri Motorin
Juergen Burhenne
Roman Teimer
Kaloian Koynov
Sophie Willnow
Elmar Weinhold
Mark Helm

Autoren-URL

https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000288800700033&DestLinkType=FullRecord&DestApp=WOS_CPL

DOI

10.1093/nar/gkq825

eISSN

1362-4962

Externe Identifier

Clarivate Analytics Document Solution ID: 740NE
PubMed Identifier: 21037259

ISSN

0305-1048

Ausgabe der Veröffentlichung

5

Zeitschrift

NUCLEIC ACIDS RESEARCH

Paginierung

1943 - 1952

Datum der Veröffentlichung

2011

Status

Published

Titel

Expanding the chemical scope of RNA:methyltransferases to site-specific alkynylation of RNA for click labeling

Sub types

Article

Ausgabe der Zeitschrift

39

Datenquelle: Web of Science (Lite)

Autoren

Y Motorin
J Burhenne
R Teimer
K Koynov
S Willnow
E Weinhold
M Helm

DOI

10.1093/nar/gkq825

eISSN

1362-4962

ISSN

0305-1048

Ausgabe der Veröffentlichung

5

Zeitschrift

Nucleic Acids Research

Sprache

en

Online publication date

2010

Paginierung

1943 - 1952

Datum der Veröffentlichung

2011

Status

Published

Herausgeber

Oxford University Press (OUP)

Herausgeber URL

http://dx.doi.org/10.1093/nar/gkq825

Datum der Datenerfassung

2020

Titel

Expanding the chemical scope of RNA:methyltransferases to site-specific alkynylation of RNA for click labeling

Ausgabe der Zeitschrift

39

Datenquelle: Crossref

Abstract

This work identifies the combination of enzymatic transfer and click labeling as an efficient method for the site-specific tagging of RNA molecules for biophysical studies. A double-activated analog of the ubiquitous co-substrate S-adenosyl-l-methionine was employed to enzymatically transfer a five carbon chain containing a terminal alkynyl moiety onto RNA. The tRNA:methyltransferase Trm1 transferred the extended alkynyl moiety to its natural target, the N2 of guanosine 26 in tRNA(Phe). LC/MS and LC/MS/MS techniques were used to detect and characterize the modified nucleoside as well as its cycloaddition product with a fluorescent azide. The latter resulted from a labeling reaction via Cu(I)-catalyzed azide-alkyne 1,3-cycloaddition click chemistry, producing site-specifically labeled RNA whose suitability for single molecule fluorescence experiments was verified in fluorescence correlation spectroscopy experiments.

Addresses

Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany.

Autoren

Yuri Motorin
Jürgen Burhenne
Roman Teimer
Kaloian Koynov
Sophie Willnow
Elmar Weinhold
Mark Helm

DOI

10.1093/nar/gkq825

eISSN

1362-4962

Externe Identifier

PubMed Identifier: 21037259
PubMed Central ID: PMC3061074

Open access

true

ISSN

0305-1048

Ausgabe der Veröffentlichung

5

Zeitschrift

Nucleic acids research

Schlüsselwörter

Organic Chemicals
Alkynes
tRNA Methyltransferases
S-Adenosylmethionine
RNA, Transfer, Phe
Fluorescent Dyes
Spectrometry, Fluorescence
Base Sequence
Molecular Sequence Data
Click Chemistry

Sprache

eng

Medium

Print-Electronic

Online publication date

2010

Open access status

Open Access

Paginierung

1943 - 1952

Datum der Veröffentlichung

2011

Status

Published

Publisher licence

CC BY-NC

Datum der Datenerfassung

2010

Titel

Expanding the chemical scope of RNA:methyltransferases to site-specific alkynylation of RNA for click labeling.

Sub types

Research Support, Non-U.S. Gov't
research-article
Journal Article

Ausgabe der Zeitschrift

39

Files

https://academic.oup.com/nar/article-pdf/39/5/1943/7168134/gkq825.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21037259/pdf/?tool=EBI https://europepmc.org/articles/PMC3061074?pdf=render

Datenquelle: Europe PubMed Central

Abstract

This work identifies the combination of enzymatic transfer and click labeling as an efficient method for the site-specific tagging of RNA molecules for biophysical studies. A double-activated analog of the ubiquitous co-substrate S-adenosyl-l-methionine was employed to enzymatically transfer a five carbon chain containing a terminal alkynyl moiety onto RNA. The tRNA:methyltransferase Trm1 transferred the extended alkynyl moiety to its natural target, the N2 of guanosine 26 in tRNA(Phe). LC/MS and LC/MS/MS techniques were used to detect and characterize the modified nucleoside as well as its cycloaddition product with a fluorescent azide. The latter resulted from a labeling reaction via Cu(I)-catalyzed azide-alkyne 1,3-cycloaddition click chemistry, producing site-specifically labeled RNA whose suitability for single molecule fluorescence experiments was verified in fluorescence correlation spectroscopy experiments.

Autoren

Yuri Motorin
Jürgen Burhenne
Roman Teimer
Kaloian Koynov
Sophie Willnow
Elmar Weinhold
Mark Helm

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/21037259

DOI

10.1093/nar/gkq825

eISSN

1362-4962

Externe Identifier

PubMed Central ID: PMC3061074

Ausgabe der Veröffentlichung

5

Zeitschrift

Nucleic Acids Res

Schlüsselwörter

Alkynes
Base Sequence
Click Chemistry
Fluorescent Dyes
Molecular Sequence Data
Organic Chemicals
RNA, Transfer, Phe
S-Adenosylmethionine
Spectrometry, Fluorescence
tRNA Methyltransferases

Sprache

eng

Country

England

Paginierung

1943 - 1952

PII

gkq825

Datum der Veröffentlichung

2011

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2011

Titel

Expanding the chemical scope of RNA:methyltransferases to site-specific alkynylation of RNA for click labeling.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

39

Datenquelle: PubMed

Expanding the chemical scope of RNA : methyltransferases to site-specific alkynylation of RNA for click labeling

Files

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